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Stress Adaptation and

Implications in Foods
DEFINITIONS
 Stress, as used in this chapter, refers to any
deleterious factor or condition that adversely
affects microbial growth or survival. According
to this practical definition, many food
processing treatments are considered
stresses.
Interrelations among physiological states of
microbial cell to different stresses .
Stresses to m.o during food production
and processing include:

 1. Physical treatments such as heat,


pressure, electric pulses, ultrasonic
waves, light/radiation, and osmotic shock
 2. Addition of chemicals such as acids,
salts, and oxidants
 3. Biological stresses, e.g., competition,
microbial metabolites and antagonism
Stress Response
 Production of proteins that repair damage,
maintain the cell, or eliminate the stress agent
 2. Transient increase in resistance or tolerance
to deleterious factors
 3. Cell transformation to a dormant state, i.e.,
spore formation or passage to the viable-but-
not-culturable state
 4. Evasion of host organism defenses
 5. Adaptive mutations
Adaptation
 When microorganisms are stressed, an
adaptive or protective response may follow.
 Response to stress, in this case, increases the
organism’s tolerance to the same or to a
different type of stress. This phenomenon is
occasionally described as adaptive response,
induced tolerance, habituation, acclimatization
or stress hardening. Stress adaptation and
stress adaptive response will be used
interchangeably in this chapter.
Tolerance
Tolerance to a deleterious factor (e.g., low pH) refers to a
microorganism’s ability to survive a stress. Each
microorganism has an inherent tolerance level to a
particular stress, but a transient or adaptive tolerance
may also be induced. For example, lactic acid bacteria
are inherently more acid tolerant than many other
bacteria, yet they can become even more acid tolerant
after acid adaptation. Resistance and tolerance have
similar meanings; these terms will be used
interchangeably in this chapter.The employment of
molecular genetic methods results in many changes
in the taxonomy of bacteria, and many of the new
taxa have been created.
Injury

 Damage to cellular components by stresses may


impair the ability of microorganisms to multiply or
may sensitize the cells to mildly deleterious
factors. These changes are commonly described
as injury. Injury is most noticeable when stress-
exposed cells become sensitive to selective
agents that healthy cells readily survive. The
relationship between cell injury and stress
adaptation has not been well characterized,
BACTERIAL TAXONOMY

 Morphological and biochemical


characteristics
 Cell wall analysis
 Serological profiles
 Cellular fatty acid profiles
BACTERIAL TAXONOMY

rRNA Analyses:
Taxonomic information can be obtained from RNA in the
production of nucleotide catalogues and the determination
of RNA sequence similarities:
 the prokaryotic ribosome is a 70S (Svedberg) unit,
which is composed of two separate functional subunits:
5OS and 30S.
 50S subunit is composed of 23 S and 5 S RNA in
addition to about 34 proteins.
 30S subunit is composed of 16S RNA plus about 21
proteins.
BACTERIAL TAXONOMY

rRNA Analyses:
The 16S subunit is highly conserved and is
considered to be an excellent chronometer of
bacteria over time, and by use of reverse
transcriptase:
 16S rRNA can be sequenced to produce long
stretches(95% ) to allow for the determination of precise
phylogenetic relationships
 To sequence 16S rRNA, a single-stranded DNA copy
is made by use of reverse transcriptase with the RNA as
template
BACTERIAL TAXONOMY

rRNA Analyses:
 When the single stranded DNA is made in the
presence of di-deoxynucleotides, DNA
fragments of various sizes result that can be
sequenced by the Sanger method
 From the DNA sequences, the template 16S
rRNA sequence can be deduced.
 Through studies of 16S rRNA sequences, three
kingdoms of life-forms: Eukaryotes, Archaebacteria,
and Prokaryotes were established.
BACTERIAL TAXONOMY

rRNA Analyses:
 with the bacteria of importance in foods being
eubacteria. Sequence similarities of 16S rRNA are
widely employed
 Libraries of eubacterial 5 S rRNA sequences also
exist, but they are fewer than for 16S.
 Nucleotide catalogs of 16S rRNA have been prepared
for a number of organisms
 The sequencing of 16S rRNA by reverse transcriptase
is preferred to oligonucleotide cataloging, as longer
stretches of rRNA can be sequenced.
BACTERIAL TAXONOMY

Analysis of DNA:
 The mol% G + C of bacterial DNA has been employed in bacterial
taxonomy for several decades
 Its use in combination with 16S and 5 S rRNA sequence data
makes it more meaningful
 When two organisms differ in G + C content by more than 10%,
they have few base sequences in common
 DNA-DNA or DNA-RNA hybridization has been employed for
some time, this technique continues to be of great value in bacterial
systematic
 The ideal reference system for bacterial taxonomy would be the
complete DNA sequence of an organism
BACTERIAL TAXONOMY

Analysis of DNA:
 Bacterial species can be defined in phylogenetic terms by use of
DNA-DNA hybridization results
 When DNA-DNA hybridization is employed, phenotypic
characteristics are not allowed to override except in exceptional
cases
 Although a genus is more difficult to define phylogenetically, 20%
sequence similarity is considered to be the minimum level of
DNA-DNA homology
 Even if there is no a satisfactory phylogenetic definition of a
bacterial genus, the continued application of nucleic acid
techniques, along with some of the other methods listed above,
should lead ultimately to a phylogenetically based system of
bacterial systematics.
COMMON FOOD-BORNE BACTERIA

 Some of the important genera known to


occur in foods are listed below in
alphabetical order.

 Some are desirable in certain foods;


Others bring about spoilage or cause
gastroenteritis.
COMMON FOOD-BORNE BACTERIA

Acinetobacter (akinetos, unable to move):


 They are gram-negative rods.
 They are strict aerobes, do not reduce nitrates.
 Although rod-shaped cells are formed in
young cultures, old cultures contain many
Coccoid-shaped cells.
 They are widely distributed in soils and waters
and may be found on many foods.
 The mol% G + C content of DNA for the genus
is 39-47.
COMMON FOOD-BORNE BACTERIA

Aeromonas (gas producing):


 These are typically aquatic gram-negative rods
 They produce copious quantities of gas from
those sugars fermented.
 They are normal inhabitants of the intestines
of fish, and some are fish pathogens.
 The mol% G + C content of DNA is 57-65.
COMMON FOOD-BORNE BACTERIA

Alcaligenes (alkali producers):


 Although gram negative, these organisms
sometimes stain gram positive.
 They are rods that do not, as the generic name
suggests, ferment sugars but instead produce alkaline
reactions, especially in litmus milk
 Nonpigmented, they are widely distributed in nature in
decomposing matter of all types
 Raw milk, poultry products, and faecal matter are
common sources.
 The mol% G + C content of DNA is 58-70
COMMON FOOD-BORNE BACTERIA

Alteromonas (anothermonad):
 These are marine and coastal water
inhabitants that are found in and on
seafoods.
 All species require seawater salinity for
growth.
 They are gram-negative motile rods that
are strict aerobes.
COMMON FOOD-BORNE BACTERIA

Arcobacter (arcus, bow):


 They are gram-negative curved or S-shaped
rods
 they can grow at 15C and are aero tolerant.
 They are found in poultry, raw milk, shellfish,
and water; and in cattle and swine products.
 These oxidative and catalase-positive
organisms cause abortion and enteritis in
some animals
COMMON FOOD-BORNE BACTERIA

Bacillus:
are gram-positive spore-forming rods that are aerobes
 Although most are mesophiles, psychrotrophs and
thermophiles exist.
 The genus contains only two pathogens: B. anthracis
(cause of anthrax) and B. cereus
 Although most strains of the latter are non-pathogens,
some cause food-borne gastroenteritis
 The mol% G + C of 51.6-60.3
 Grow as low as about 35°C to 70°C, and over the pH
range of 2 to 6.
COMMON FOOD-BORNE BACTERIA

Brochothrix (brochos, loop; thrix, thread):


 These gram-positive non-sporeforming rods
 They are common on processed meats and on
fresh and processed meats that are stored in
gas-impermeable packages at refrigerator
temperatures.
 They are rhamnose and hippurate positive.
 The mol% G + C content of DNA is 36.
 They do not grow at 37°C.
COMMON FOOD-BORNE BACTERIA

Campylobacter (campylo, curved):


 These are gram-negative.
 Spirally curved rods.
 They are microaerophilic to anaerobic
 The mol% G + C content of DNA is 30-
35.
COMMON FOOD-BORNE BACTERIA

Carnobacterium (flesh-meat bacteria):


 These are grampositive.
 Catalase-negative rods.
 They are heterofermentative.
 Most grow at 0C and none at 45C.
 Gas is produced from glucose by some
species.
 The mol% G + C for the genus is 33.0-37.2.
COMMON FOOD-BORNE BACTERIA

Citrobacter:
 These enteric bacteria are slow lactose-
fermenting
 Gram-negative rods
 Can use citrate as the sole carbon source
 Most prevalent species in foods
 The mol% G + C content of DNA is 50-52.
COMMON FOOD-BORNE BACTERIA

Clostridium (closter, a spindle):


 These anaerobic poreforming rods are widely
distributed in nature.
 The genus contains many species, some of
which cause disease in humans.
 Mesotrophic, psychrotrophic, and thermophilic
species/strains exist.
COMMON FOOD-BORNE BACTERIA

Corynebacterium (coryne, club):


 The true coryneform genera of gram-positive,
rod-shaped bacteria.
 They are sometimes involved in the spoilage
of vegetable and meat products
 Most are mesotrophs, although psychrotrophs
are known.
 C. diphtheriae, causes diphtheria in humans.
 The mol% G + C content of DNA is 51-63.
COMMON FOOD-BORNE BACTERIA

Enterobacter:
 These enteric gram-negative bacteria.
 They are typical of other
Enterobacteriaceae relative to growth
requirements, although they are not
generally adapted to the gastrointestinal
tract.
COMMON FOOD-BORNE BACTERIA

Enterococcus:
 They are grampositive ovoid cells that
occur singly, in pairs, or in short chains.
 They have phylogenetic relationship to
other lactic acid bacteria.
COMMON FOOD-BORNE BACTERIA

Erwinia:
 These are gram-negative enteric rods.
 They are especially associated with
plants, where they cause bacterial soft
rot .
The mol% G + C content of DNA is 53.6-
54.1.
COMMON FOOD-BORNE BACTERIA

Escherichia:

 This is the most widely studied genus of all


bacteria.
 They cause food-borne gastroenteritis
 E. coli can be used as an indicator of food
safety.
COMMON FOOD-BORNE BACTERIA

Flavobacterium:
 These are gram-negative rods.
 They are characterized by their production of
yellow to red pigments on agar and by their
association with plants.
 Some are mesotrophs, and others are
psychrotrophs.
 They participate in the spoilage of refrigerated
meats and vegetables.
 Some of the new genera contain fish
pathogens and some are halophiles.
COMMON FOOD-BORNE BACTERIA

Lactobacillus:
 They are gram-positive.
 catalase-negative rods that often occur in long chains.
 Although those in foods are typically microaerophilic,
many true anaerobic strains exist, especially in human
stools and the rumen
 They occur on most vegetables and dairy products,
they also recovered from apple and pear mashes.
 They grows at pH 2.8 in 12-16% ethanol.
COMMON FOOD-BORNE BACTERIA

Lactococcus:
 They are gram-positive, nonmotile.
 They are catalase-negative spherical or ovoid
cells that occur singly, in pairs, or as chains.
 They grow at 10C but not at 45C.
 Most strains react with group N antisera.
 L-Lactic acid is the predominant end product
of fermentation.
COMMON FOOD-BORNE BACTERIA

Listeria:

 They are gram-positive, nonsporing rods.


 They have identical cell walls, fatty acid, and
cytochrome composition.
COMMON FOOD-BORNE BACTERIA

Micrococcus:
 These cocci are gram positive and catalase
positive.
 Some produce pink to orange-red to red
pigments.
 Most can grow in the presence of high levels
of NaCl.
 Most are mesotrophs, although psychrotrophic
species/strains are known.
 The mol% G + C content of DNA of 69-76.
COMMON FOOD-BORNE BACTERIA

Moraxella:
 These short gram-negative rods.
 Sensitive to penicillin.
 Oxidase positive.
 They having a mol% G + C DNA content
of 40-46.
COMMON FOOD-BORNE BACTERIA

Pediococcus (coccus growing in one plane):


 They are homo-fermentative cocci.
 They are lactic acid bacteria that exist in
pairs and tetrads resulting from cell
division in two planes.
 Their mol% G + C content of DNA is
34.4.
 They can grow in 18% NaCl.
COMMON FOOD-BORNE BACTERIA

Pseudomonas (false monad):


 These are gram-negative rods.
 Constitute the largest genus of bacteria that exists in
fresh foods.
 The mol% G + C content of their DNA of 58-70.
 This is a heterogeneous group.
 They are typical of soil and water bacteria.
 Widely distributed among foods, especially
vegetables, meat, poultry, and seafood products.
 Many species and strains are psychrotrophic.
 Some produce water-soluble, blue-green pigments.
COMMON FOOD-BORNE BACTERIA

Psychrobacter:
 They are non-motile gram-negative rods, plump
coccobacilli that occur often in pairs.
 They do not ferment glucose.
 Growth occurs in 6.5% NaCl and at 1°C, but not at
35°C
 They hydrolyze Tween 80, and most are egg-yolk
positive (
 They are sensitive to penicillin and utilize 7-
aminovalerate
 They unable to utilize glycerol or fructose.
 They are common on meats, poultry, fish, & in waters.
COMMON FOOD-BORNE BACTERIA

Salmonella:
 All members of this genus of gram-negative
enteric bacteria.
 They are considered to be human pathogens.
 The mol% G + C content of DNA is 50-53.
COMMON FOOD-BORNE BACTERIA

Shigella:
 All members of this genus are presumed
to be human enteropathogens.
COMMON FOOD-BORNE BACTERIA

Staphylococcus (grapelike coccus):

 These are gram-positive.


 Catalase-positive cocci.
 They include S. aureus, which causes several
disease syndromes in humans, including food-
borne gastroenteritis.
COMMON FOOD-BORNE BACTERIA

Vibrio:

 These gram-negative straight or curved rods.


 Several species cause gastroenteritis and
other human illness.
 The mol% G + C content of DNA is 38-51.
COMMON FOOD-BORNE BACTERIA

Yersinia:
 This genus includes the agent of human
plague, Y.pestis
 They cause food-borne gastroenteritis, Y.
enterocolitica
 The mol% G + C content of DNA is 45.8-46.8.
The sorbose-positive biogroup 3A strains have
been elevated as Y. mollaretti
 The sorbose-negative strains as Y bercovieri.
COMMON GENERA OF
FOOD-BORNE MOLDS

 Moulds are filamentous fungi that grow


in the form of a tangled mass that
spreads rapidly and may cover several
inches of area in few days.
 The total of the mass or any large
portion of it is referred to as mycelium.
 Mycelium is composed of branches or
filaments referred to as hyphae.
COMMON GENERA OF
FOOD-BORNE MOLDS

 Those of greatest importance in foods


multiply by ascospores, zygospores, or
conidia.
 The ascospores of some genera are
notable for their extreme degrees of heat
resistance.
The Taxonomy of Molds

 No radical changes in the systematics of


foodborne fungi
 The most notable changes involve the
discovery of the sexual or perfect states
of some well-known genera and species.
 The ascomycete state is believed to be
the more important reproductive state of
a fungus, and this state is referred to as
the teleomorph.
The Taxonomy of Molds

 The species name given to a teleomorph


takes precedence over that for the
anamorph, the imperfect or conidial
state.
 Holomorph indicates that both states are
known, but the teleomorph name is
used.
 The taxonomic positions of the genera
described are summarized below: